3.  The presence or absence of light is important.  Nitrification is most rapid in darkness; and in the case of solutions, exposure to strong light may cause nitrification to cease altogether.

4.  The presence of oxygen is of course essential.  A thin layer of solution will nitrify sooner than a deep layer, owing to the larger proportion of oxygen available.  The influence of depth of fluid is most conspicuous in the case of strong solutions.

5.  The quantity of nitrifying organism present has also a marked effect.  A solution seeded with a very small amount of organism will for a long time exhibit no nitrification, the organism being (unlike some other bacteria) of very slow growth.  A solution receiving an abundant supply of the ferment will exhibit speedy nitrification, and strong solutions may by this means be successfully nitrified, which with small seedings would prove very refractory.  The speedy nitrification which occurs in soil (far more speedy than in experiments in solutions under any conditions yet tried) is probably owing to the great mass of nitrifying organisms which soil contains, and to the thinness of the liquid layer which covers the soil particles.

6.  The rapidity of nitrification also depends on the degree of alkalinity of the solution.  Nitrification will not take place in an acid solution; it is essential that some base should be present with which the nitric acid may combine; when all available base is used up, nitrification ceases.

It appeared of interest to ascertain to what extent nitrification would proceed in a dilute solution of urine without the addition of any substance save the nitrifying ferment.  As urea is converted into ammonium carbonate in the first stage of the action of the ferment, a supply of salifiable base would at first be present, but would gradually be consumed.  The result of the experiment showed that only one-half the quantity of nitric acid was formed in the simple urine solution as in similar solutions containing calcium and sodium carbonate.  The nitrification of the urine had evidently proceeded until the whole of the ammonium had been changed into ammonium nitrate, and the action had then ceased.  This fact is of practical importance.  Sewage will be thoroughly nitrified only when a sufficient supply of calcium carbonate, or some other base, is available.  If, instead of calcium carbonate, a soluble alkaline salt is present, the quantity must be small, or nitrification will be seriously hindered.

Sodium carbonate begins to have a retarding influence on the commencement of nitrification when its amount exceeds 300 milligrammes per liter, and up to the present time I have been unable to produce an effective nitrification in solutions containing 1.000 gramme per liter.

Sodium hydrogen carbonate hinders far less the commencement of nitrification.

Ammonium carbonate, when above a certain amount, also prevents the commencement of nitrification.  The strongest solution in which nitrification has at present commenced contained ammonium carbonate equivalent to 368 milligrammes of nitrogen per liter.  This hinderance of nitrification by the presence of an excess of ammonium carbonate effectually prevents the nitrification of strong solutions of urine, in which, as already mentioned, ammonium carbonate is the first product of fermentation.